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Reovirus Neuropathogenesis

呼肠孤病毒神经发病机制

基本信息

批准号：
10607594
负责人：
TERENCE S. DERMODY
金额：
$ 56.55万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-22 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10607594
关键词：
3-Dimensional Adult Afferent Neurons Atomic Force Microscopy Axon Axonal Transport Binding Biophysics Brain Brain imaging CRISPR-mediated transcriptional activation Capsid Capsid Proteins Cells Cerebellum Child Clustered Regularly Interspaced Short Palindromic Repeats Cryoelectron Microscopy Cultured Cells Development Disease Double Stranded RNA Virus Dynein ATPase Electron Microscopy Encephalitis Engineering Fiber Genome Hippocampus (Brain)Human Immunocompetent Immunoglobulins In Vitro Infection Integration Host Factors Integrins Interneurons Intervention Intramuscular Knockout Mice Knowledge Malignant Neoplasms Mammals Mediating Morbidity - disease rate Motor Mus Mutagenesis Nervous system structure Neuraxis Neurons Neuropathogenesis Neurotropism Newborn Infant Oncolytic Pattern Peripheral Pharmacology Population Process Proteins Reovirus Reovirus Infections Research Resolution Role Route Sensory Receptors Series Serotyping Structure Synapses System Testing Thalamic structure Tissues Tropism Viral Viral Encephalitis Viral Pathogenesis Virulence Virus Work blind cell type clinical development experimental study fast axonal transport in vivo mortality mutant nervous system disorder neural circuit neuronal cell body neurotropic neurotropic virus neurovirulence new therapeutic target oncolytic vector particle receptor receptor function reconstruction recruit relating to nervous system reverse genetics screening targeted treatment transmission process uptake virus host interaction

项目摘要

Viral encephalitis is an important cause of morbidity and mortality in children and adults. Key knowledge gaps about the pathogenesis of viral encephalitis include how neurotropic viruses target the central nervous system (CNS), internalize into neurons, and disseminate in the brain. The proposed research uses reovirus, a genetically tractable double-stranded RNA virus that causes encephalitis and shows promise as an oncolytic agent, to dissect mechanisms of viral tropism, cell entry, and spread in the CNS. Reovirus strains display serotype-specific patterns of systemic dissemination and neurologic disease. Serotype 3 (T3) reoviruses are exquisitely neurotropic, internalize into neurons using macropinocytosis, and spread intracellularly by fast axonal transport. Reovirus uses β1 integrin to internalize into some types of cells, but its function in neural cell entry is unknown. Other previously identified reovirus receptors are dispensable for reovirus infection in the murine CNS. We recently identified a new reovirus receptor, paired immunoglobulin-like receptor B (PirB), that allows serotype-independent reovirus infection of cultured cells but is required for efficient T3 infection of neurons and full neurovirulence. Three integrated specific aims are proposed to enhance knowledge of reovirus neuropathogenesis that may have broader applications to other neurotropic viruses. In Specific Aim 1, functions of host receptors in reovirus neuropathogenesis will be determined. Biophysical interactions between PirB and reovirus will be assessed using cryo-electron microscopy, atomic-force microscopy, and structure- guided mutagenesis. The function of PirB in reovirus disease will be elucidated by comparing infection and virulence in wild-type (WT) and PirB-null mice. T3 reovirus-specific receptors that couple with PirB for neurotropism will be identified using CRISPR activation screening. In Specific Aim 2, mechanisms of reovirus entry into CNS neurons will be defined. The functions of PirB and β1 integrin in reovirus neural entry will be determined using mutant receptors and receptor-blind viruses. Receptor-dependent control of macropinocytosis will be elucidated using super-resolution, live-cell, and electron microscopy. Mechanisms of dynein motor recruitment for reovirus transport within axons will be dissected by investigating interactions between WT and mutant reovirus receptors and dynein subunits. In Specific Aim 3, the basis of reovirus dissemination in the CNS will be elucidated. The role of host receptors in reovirus neural transit will be defined using whole-brain imaging and three-dimensional reconstruction to trace reovirus infection and neural spread in WT and receptor- null mice. Mechanisms of reovirus transsynaptic transmission will be elucidated using in vitro and in vivo neural circuits. The effect of altering synaptic activity on reovirus neural transit will be tested using pharmacologic intervention and chemogenetically altered mice. Taken together, these studies will define mechanisms used by neurotropic reovirus to infect and disseminate in the brain and may promote further development of reovirus oncolytics that selectively target the nervous system.

病毒性脑炎是儿童和成人发病和死亡的重要原因。关键知识缺口